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Felis Concolor Coryi) J. Helminthol. Soc. Wash. 64(1), 1997 pp. 137-141 Microfilariae in the Free-Ranging Florida Panther (Felis concolor coryi) MARNIE G. LAMM,' MELODY E. RoELKE,2-5 ELLIS C. GREiNER,3-6 AND CHRISTINE K. STEiBLE4 1 College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610 (e-mail: [email protected]), 2 Florida Game and Fresh Water Fish Commission, Gainesville, Florida 32610, 3 Department of Infectious Diseases, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610 (e-mail: [email protected].), and 4 Department of Statistics, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Florida 32611 ABSTRACT: Blood samples from Florida panthers (Felis concolor coryi) collected from 1986 to 1993 during the months of December through May were screened for the presence of microfilariae (mff) by the Difil® filter test. Thirty-five of 47 (74.5%) panthers older than 2 yr of age were positive with microfilaremias ranging from 10 to 7,380 mff/ml of whole blood. No panthers that were 6 mo of age or less (n = 10) were microfilariae- positive, and only 20% of the panthers in the 1-yr class (n = 5) were positive. A representative number of microfilariae (n = 40) from each of 7 freshly collected positive blood samples was measured and morphological characteristics were noted. The average length of microfilariae processed by the modified Knott's technique was 320 /Am (273-370 /Am) with a width of 4-5 /Am. Of the 280 microfilariae measured, 202 (72.14%) had tapered heads and straight tails with an average length of 319 /im (276-368 /Am), 61 (21.79%) had blunt heads and straight tails and averaged 323 /urn (274-366 /Am), 16 (5.71%) had tapered heads and button-hooked tails with an average length of 320 /Am (290-368 /am), and 1 (0.35%) had a blunt head and button-hooked tail and measured 320 /Am. The finding of no significant difference (P > 0.05) between length measurements due to differences in head and tail shape leads us to believe that all microfilariae were of 1 species. Based on micro- filarial length measurements, review of necropsy reports, and comparison with bobcat microfilariae, the most likely filarial species infecting the Florida panther is Dirofilaria striata (Molin, 1858). KEY WORDS: Florida panther, Felis concolor coryi, Dirofilaria striata, microfilariae, morphology, prevalence, bobcat, Lynx rufus. Filarial nematodes have been reported in a been found in the ocelot (Felis pardalis), the number of exotic felid species. Dirofilaria re- margay (Felis tigrina) (Anderson and Diaz-Un- pens has been reported in the lion (Panthera leo) gria, 1959), and the bobcat (Orihel and Ash, (Nelson et al., 1962). Dirofilaria immitis has 1964; Miller and Harkema, 1968; Roelke, un- been found in jaguars (Felis onca), tigers (Felis publ. 1985). Forrester et al. (1985) reported sondiacus and Felis tigris), wild cats (Felis adult D. striata in mff+ adult Florida panthers. bangsi costariensis), jagouarundi (Felis yago- The Florida panther is an endangered subspe- uarundi) (Otto, 1974), a bengal tiger (Panthera cies of cougar that inhabits pockets of habitat in tigris) (Kennedy and Patton, 1981), and bobcats the Big Cypress and Everglades ecosystems of (Lynx rufus) from Florida (Levine, 1980; For- southern Florida (U.S.A.). A population of less rester, 1992). Of 22 free-ranging mountain lions than 40 adult animals remains in the wild (Bel- (Felis concolor) from California, 1 was seropos- den, 1986). Since veterinary involvement in the itive for antibodies against the somatic antigen Florida Panther Recovery Program began in of D. immitis yet was seronegative for cuticular 1983, an intensive health evaluation and moni- antigen and no circulating microfilariae (mff) toring program has been in effect. The protocol were found in the whole blood (Paul-Murphy et includes the collection, screening, and storage of al., 1994). Dirofilaria striata was first reported various biological samples each time an animal in Brazilian pumas (Felis concolor and F. ma- is immobilized. In the past, the presence of mff croura) (Raillet and Henry, 1911) and since has has been noted either by buffy coat analysis, by membrane filtration, or in reports from clinical laboratories. Adult D. striata were found in 4 5 Present address: Tanzania National Parks, Arusha, Tanzania, Africa. out of the 7 panthers examined from 1973 to 6 Corresponding author. 1983, 3 of which were mff+ (Forrester et al., 137 Copyright © 2011, The Helminthological Society of Washington 138 JOURNAL OF THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON, 64(1), JAN 1997 1985). It was assumed that all microfilariae from high numbers of mff were chosen for morphological these animals were representative of D. striata study of individual microfilariae. All blood samples were kept in EDTA-coated blood tubes for the 2-8 hr because no other adult filariids were detected. between time of collection and time of microfilarial Adult D. striata were found in 5 additional analysis. A 1-ml aliquot was processed using a modi- microfilaremic panthers at necropsy (Roelke, fied Knott's procedure (Knott, 1939). Microfilariae Nayer, and Vickery, unpub. data 1985). Accord- were measured immediately after processing in order to minimize possible effects on length or width of mi- ing to necropsy reports, 1-3 adult filariids were crofilariae due to storage in 2% formalin. Measure- present singularly in the fascia between muscle ments and morphological assessments were made with bundles in the distal extremities. No reports of a calibrated ocular micrometer at a magnification of any filarial species other than D. striata were X400. Length, head shape, tail shape, and body shape found in review of Florida panther necropsy re- were recorded for 40 mff/sample. Head shape was con- sidered tapered if the width decreased upon successive ports (n = 41) from 1980 to September 1994. measurements anteriorly (Fig. 1); if the sides remained This study was undertaken to determine the parallel, the head was categorized as blunt (Fig. 2). prevalence and microfilaremia of Dirofilaria sp. Although many posterior ends were curved, only those in the Florida panther population and to char- that had a distinct hook at the tip were labeled as such (Fig. 1,4). acterize and measure the mff in order to confirm Data were analyzed using SAS (SAS Institute Inc., the identity of the species of filariid present. 1989). Comparisons of total lengths of the microfilar- iae were made between panthers and bobcats and be- Materials and Methods tween morphological types using ANOVA with a split- plot design where felid type was the whole-plot factor From 1986 to 1993, a total of 83 blood samples and morphological type was the sub-plot factor. Three representing 47 individual panthers and 3 bobcats was morphological types (tapered head and straight tail, screened for the presence of mff. Samples were col- blunt head and straight tail, and tapered head and but- lected during the field capture season in the months of ton-hooked tail) were included in this analysis. Micro- December through May from free-ranging panthers in filariae with blunt head and button-hooked tails were the Big Cypress and Everglades ecosystems. All ani- not included due to low prevalence (1 found in pan- mals were anesthetized with ketamine hydrochloride thers and 3 found in bobcats). Confidence limits were (Ketastat®, Bristol Laboratories, Syracuse, New York, not calculated for percentage of prevalence of mff in U.S.A.) or a combination of tiletamine/zolazepam (Tel- panthers because the number examined represented azol®, A. H. Robins Co., Richmond, Virginia, U.S.A.). nearly all of the extant population of panthers, thus Blood was drawn prior to administration of fluids, vac- rendering confidence limit calculations moot. cinations, and prophylactic injections of Ivermectin (Ivomec®, Merck & Company, Incorporated, Rahway, New Jersey, U.S.A.) at a dose of 200 meg/kg. Blood Results and Discussion samples were taken directly from the saphenous vein Thirty-five (74.5%) of 47 adult (>2 yr of age) into sterile 3-ml tubes containing ethylenediaminetet- Florida panthers sampled from 1986 to 1993 raacetic acid (EDTA) via a vacutainer and butterfly apparatus. The sample was kept cool in an insulated were mff+ at some point in their lives and all container for the 2-8-hr transition from field to lab. except for 2, which had low counts of 20 and One milliliter of EDTA preserved whole blood from 330 mff/ml, remained positive on subsequent each of the 17 samples collected in 1992 and 5 sam- tests. Because panthers were usually sampled at ples from 1993 was processed using the Difil® (EV- SCO Pharmaceuticals, Buena, New Jersey, U.S.A.) 2-yr intervals, the average age when animals be- procedure described by Howland and Todd (1977). In come mff+ is not precisely known. None of the addition, 58 whole blood samples preserved in EDTA 5 panthers tested at 6 mo of age were mff+, 2 that had been collected from free-ranging panthers of 10 (20%) tested positive in the 1-yr-old class, from 1986 to 1991 and stored in a 1:10 ratio of blood and 15 of 23 (65%) of panthers were test posi- (1 ml) to 2% formalin (9 ml) at 4°C were screened for mff. From the 10-ml sample, 5 ml were passed through tive at 2-4 yr of age. Of the 23 panthers that a Difil filter. Numbers of mff were determined for all were 10 yr of age or older, 22 (96%) were mff+. positive samples (except FP#5 1 in which high numbers Counts ranged from 10 to 7,380 mff/ml of of mff/field made accurate counting impossible) by whole blood. Microfilarial counts showed fluc- counting all individual mff on the filter at a viewing magnification of X200 to obtain a mff/1 ml value.
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